Document Type : Original Article


1 Department of Range Management, Faculty of Natural Resources, Sari Agricultural Sciences and Natural Resources University, Sari, Iran

2 Department of Forestry, Faculty of Natural Resources, Sari Agricultural Sciences and Natural Resources University, Sari, Iran

3 Department of Watershed Management, Faculty of Natural Resources, Sari Agricultural Sciences and Natural Resources UniversityT Sari, Iran

4 Department of Soil Science Engineering, Faculty of Agricultural Engineering and Technology, College of Agriculture and Natural Resources, Karaj, Iran


Introduction: Mining activities causes substantial damage to the environment worldwide. Abandoned mine wastes from mining activities limit the growth of plants due to unfavorable conditions and the concentration of a wide range of heavy metals. Identifying plant species growing on mine waste and assessing their seed germinations are important for vegetation restoration on mining areas. This study aimed at assessing the germination characteristics of Artemisia absinthium, Lepidium draba, and Silybum marianum naturally growing on coal mine wastes in Mazandaran Province under different concentrations of copper (Cu), lead (Pb), and cadmium (Cd).
Material and methods: Seeds were collected from plants growing in coal wastes during 2 years. Completely randomized design was conducted with three replicates. Treatments were CuSO4 and Pb(NO3)2 at 0 (control), 50, 100, 200, and 300 mg/L, and Cd(NO3)2 at 0 (control), 5, 10, 20, and 30 mg/L. In each replicate 20 seeds were placed in a Petri dish containing a layer of filter paper. Then treatments were applied and Petri dishes were taken to a germinator under controlled temperature, moisture, and light. Germinated seeds were counted daily and then germination percentage, germination rate, allometric coefficient, seed vigor index, phytotoxicity percentage and seed tolerance index were calculated.
Results and discussion: The results showed that increasing the concentration of lead, copper, and cadmium significantly affected the seed germination of studied plants. S. marianum had better germination percentage and rate in lead, and copper while germination of A. absinthium, L. draba, was better in cadmium, and copper, respectively. The greatest reduction in germination percentage was found for A. absinthium in 20 and 30 mg/L Cd. As the concentration of metals increased, significant reduction in germination percentage and rate for A. absinthium started in lower concentrations than other species. Root and shoot length of all species was significantly reduced even in low concentration of metals and S. marianum had greater root and shoot length under all treatments. Reduction in seed vigor index was greater under Cu than that in Pb and Cd. High tolerance index was found for L. draba in response to  Cd and for S. marianum under the stress of  Pb and  Cu. In all treatments, an increase in metal concentration significantly increased the phytotoxicity index. Under Cu and Pb S. marianum showed the least phytotoxicity index, while the lowest phytotoxicity index was found for L. draba under Cd stress. Cu and Cd in high concentration were more toxic for A. absinthium than that for the other two species. Under similar concentrations of Cu and  Pb, all species showed reduced germination in copper stress. Cd in lower concentrations were tolerable for plants but generally it had the greatest inhibition especially on A. absinthium.
Conclusion: Overall the responses were different regarding the plant species, metal and concentrations. All three species were able to germinate at high concentrations of heavy metals. The presence of these species in coal wastes and their ability to germinate in high concentrations of heavy metals makes it possible to use these plants in future restoration and phytoremediation programs.


Abdul-Baki, A.A. and Anderson, J.D., 1973. Vigor determination in soybean seed by multiple criteria. Crop Science. 13(6), 630-633.
Abraham, K., Sridevi, R., Suresh, B. and Damodharam, T., 2013. Effect of heavy metals (Cd, Pb, Cu) on seed germination of Arachis hypogeae L. Asian Journal of Plant Science and Research. 3(1), 10-12.
Aflaki, F., Sedghi, M., Pazuki, A. and Pessarakli, M., 2017. Investigation of seed germination indices for early selection of salinity tolerant genotypes: A case study in wheat. Emirates Journal of Food and Agriculture. 29(3), 222-226.
Agraval, R.L., 2005. Seed Technology. Oxford and IBH Publishing Company, UK.
Ahmadiyan, E. and Motesharezade, B., 2016. Study heavy metals concentration changes and ecological index pollution soils around charcoal mine Geland-roud Mazandaran Province. Journal of Land Management. 3(2), 73-81. (In Persian with English abstract).
Ahsan, N., Lee, D.G., Lee, S.H., Kang, K.Y., Lee, J.J., Kim, P.J., Yoon, H.S. Kim, J.S. and Lee, B.H., 2007. Excess copper induced physiological and proteomic changes in germinating rice seeds. Chemosphere. 67(6), 1182-1193.
Ali, H., Khan, E. and Sajad, M.A., 2013. Phytoremediation of heavy metals-concepts and applications. Chemosphere. 91(7), 869-881.
Benavides, M.P., Gallego, S.M. and Tomaro, M.L., 2005. Cadmium toxicity in plants. Brazilian Journal of Plant Physiology. 17(1), 21-34.
Bezini, E., Abdelguerfi, A., Nedjimi, B., Touati, M., Adli, B. and Yabrir, B., 2019. Effect of some heavy metals on seed germination of Medicago arborea L. (Fabaceae). Agriculturae Conspectus Scientificus. 84(4), 357-364.
Blowes, D.W., Ptacek, C.J., Jambor, J.L., Weisener, C.G., Paktunc, D., Gould, W.D. and Johnson, D.B., 2014. The geochemistry of acid mine drainage. In: Holland, H.D. and Turekian, K.K. (Eds.), Treatise on Geochemistry. Elsevier, Oxford, UK. pp. 131–190.
Boi, M.E., Porceddu, M., Cappai, G., De Giudici, G. and Bacchetta, G., 2020. Effects of zinc and lead on seed germination of Helichrysum microphyllum subsp. tyrrhenicum, a metal-tolerant plant. International Journal of Environmental Science and Technology. 17, 1917–1928.
Chiapusio, G., Sanchez, A.M., Reigosa, M.J., Gonzalez, L. and Pellissier, F., 1997. Do germination indices adequately reflect allelochemical effects on the germination process? Journal of Chemical Ecology. 23(11), 2445-2453.
Chou, C.H. and Lin, H.J., 1976. Autointoxication mechanism of Oryza sativa I. Phytotoxic effects of decomposing rice residues in soil. Journal of Chemical Ecology. 2(3), 353-367.
da Silva, E., Guilherme, M.F.S., Oliveira, H.M., Araujo, L.N.C.P., Viana, Z.C.V. and Santos, V.L.C.S., 2017. Ecotoxicological effects of cadmium on the germination and initial development of Schinus terebinthifolius. Revista de Ciencias Agrarias. 40(2), 311-318.
Delgado-Caballero, M.D.R., Alarcon-Herrera, M.T., Valles-Aragon, M.C., Melgoza-Castillo, A., Ojeda-Barrios, D.L. and Leyva-Chavez, A., 2017. Germination of Bouteloua dactyloides and Cynodon dactylon in a multi-polluted soil. Sustainability. 9, 1-9.
Ebrahimi, M. and Madrid Diaz, F., 2014. Use of Festuca ovina L. in chelate assisted phytoextraction of copper contaminated soils. Journal of Rangeland Science. 4(3), 171-182. (In Persian with English abstract).
El Rasafi, T., Nouri, M., Bouda, S. and Haddioui, A., 2016. The effect of Cd, Zn and Fe on seed germination and early seedling growth of wheat and bean. Ekologia (Bratislava). 35(3), 213-223.
Farias, J.G., Antes, F.L., Nunes, P.A., Nunes, S.T., Schaich, G., Rossato, L.V., Miotto, A., Girotto, E., Tiecher, T.L., Dressler, V.L. and Nicoloso, F.T., 2013. Effects of excess copper in vineyard soils on the mineral nutrition of potato genotypes. Food and Energy Security. 2(1), 49-69.
Golubovic Curge, V., Raicevic, V., Veselinovic, M., Tabakovic-Tosic, M. and Vilotic, D., 2012. Influence of heavy metals on seed germination and growth of Picea abies L. Karst. Polish Journal of Environmental Studies. 21(2), 353-359.
Han, Y.L., Yuan, H.Y., Huang, S.Z., Guo, Z., Xia, B. and Gu, J., 2007. Cadmium tolerance and accumulation by two species of Iris. Ecotoxicology. 16, 557-563.
Hatamzadeh, A., Sharaf, A.R.N., Vafaei, M.H., Salehi, M. and Ahmadi, G., 2012. Effect of some heavy metals (Fe, Cu and Pb) on seed germination and incipient seedling growth of Festuca rubra ssp. commutate (Chewings fescue). International Journal of Agriculture and Crop Science. 4, 1068-1073.
Iqbal, M.Z. and Rahmati, K., 1992. Tolerance of Albizia lebbeck to Cu and Fe application. Ekologia. 11(4), 427-430.
Iqbal, M.Z. and Shazia, Y., 2004. Reduction of germination and seedling growth of Leucaena leucocephala caused by lead and cadmium individually and in combinations. Ekologia (Bratislava). 23(2), 162-168.
ISTA. 2017. International Rules for Seed Testing. International Seed Testing Association (ISTA), Bassersdorf.
Jamalloo, F. and Ghorbani Ghomi, T., 2015. Study of morphology and Pollen grain Cardaria draba (L.) DESV. of Brassicaceae in Iran. Journal of Environmental Science and Technology, 16(1): 647-653. (In Persian with English abstract).
Kabata-Pendias, A., 2000. Trace Elements in Soils and Plants. CRC press, Boca Raton.
Khajeh-Hosseini, M., Powell, A.A. and Bingham, I.J., 2003. The interaction between salinity stress and seed vigor during germination of soyabean seeds. Seed Science and Technology. 31(3), 715-725.
Ko, K.S., Lee, P.K. and Kong, I.C., 2012. Evaluation of the toxic effects of arsenite, chromate, cadmium, and copper using a battery of four bioassays. Applied Microbiology and Biotechnology. 95(5), 1343-1350.
Kranner, I. and Colville, L., 2011. Metals and seeds: biochemical and molecular implications and their significance for seed germination. Environmental and Experimental Botany. 72(1), 93-105.
Lashgari, N., Ghorbani, J., Zali, S.H. and Vahabzadeh, G.H., 2016. Assessing the vegetation restoration potential on coal mine waste (Case study: Karmozd Savadkoh mines, Mazandaran province). Journal of Environmental Studies. 41(76), 757-770. (In Persian with English abstract).
Lashgari, N., Ghorbani, J., Zali, S.H. and Vahabzadeh, G.H., 2017. Soil properties and level of heavy metals in coal wastes and their association with plant establishment (Case study: coal mine of Karmozd Svadkoh, Mazandaran province). Journal of Natural Environment. 69(4), 1091-1108. (In Persian with English abstract).
Lefevre, I., Marchal, G., Correal, E., Zanuzzi, A. and Lutts, S., 2009. Variation in response to heavy metals during vegetative growth in Dorycnium pentaphyllum Scop. Plant Growth Regulation. 59(1), 1-11.
Li, W., Khan, M.A., Yamaguchi, S. and Kamiya, Y., 2005. Effects of heavy metals on seed germination and early seedling growth of Arabidopsis thaliana. Plant Growth Regulation. 46(1), 45-50.
Mahmood, S., Hussain, A., Saeed, Z. and Athar, M., 2005. Germination and seedling growth of corn (Zea mays L.) under varying levels of copper and zinc. International Journal of Environmental Science and Technology. 2(3), 269-274.
Masto, R.E., George, J., Rout, T.K. and Ram L.C., 2017. Multi element exposure risk from soil and dust in a coal industrial area. Journal of Geochemical Exploration. 176, 100-107.
McKenna, D.J., Jones, K. and Hughes, K., 2002. Botanical Medicines: The Desk Reference
for Major Herbal Supplements. Haworth Press, Binghamton, New York.
Mihoub, A., Chaoui, A. and El Ferjani, E., 2005. Biochemical changes associated with cadmium and copper stress in germinating pea seeds (Pisum sativum L.). Comptes Rendus Biologies. 328(1), 33-41.
Monaci, F., Trigueros, D., Mingorance, M.D. and Rossini-Oliva, S., 2019. Phytostabilization potential of Erica australis L. and Nerium oleander L.: a comparative study in the Riotinto mining area (SW Spain). Environmental Geochemistry and Health. 42, 2345-2360.
Mozaffarian, V.A., 2008. Flora of Iran: The Family of Asteraceae, First ed., Research Institute of Forests and Rangelands, Tehran.
Namjooyan, R., Ghorbani, J., Heydari, GH.A. and Vahabadeh, G.H., 2019. Presence of rangeland plant species in the seed bank of coal waste and surrounding rangelands in Karmozd-Savad kouh and Kiasar-Sari coal mines in Mazandaran province. Journal of Range and Watershed Management. 72(2), 587-596. (In Persian with English abstract).
Peto, A., Lehotai, N., Lozano-Juste, J., Leon, J., Tari, I., Erdei, L. and Kolbert, Z., 2011. Involvement of nitric oxide and auxin in signal transduction of copper-induced morphological responses in Arabidopsis seedlings. Annals of Botany. 108(3), 449-457.
Pietrini, F., Iannelli, M.A., Pasqualini, S. and Massacci, A., 2003. Interaction of cadmium with glutathione and photosynthesis in developing leaves and chloroplasts of Phragmites australis (Cav.) Trin. ex Steudel. Plant Physiology. 133(2), 829-837.
Pinto, A.P., Mota, A.M., de Varennes, A. and Pinto, F.C., 2004. Influence of organic matter on the uptake of cadmium, zinc, copper and iron by sorghum plants. Science of the Total Environment. 326, 239–247.
Prach, K., Bartha, S., Joyse, B., Pysek, P., Van Diggelen, R. and Wiegleb, G., 2001. The role of spontaneous vegetation succession in ecosystem restoration: A perspective. Applied Vegetation Science. 4(1), 111-114.
Radha, J., Srivastava, S., Solomon, S., Shrivastava, A.K. and Chandra, A., 2010. Impact of excess zinc on growth parameters, cell division, nutrient accumulation, photosynthetic pigments and oxidative stress of sugarcane (Saccharum spp.). Acta Physiologiae Plantarum. 32(5), 979-986.
Saberi, M. and Karimian, V., 2019. Effects of chemical stimulators to development, support and resistant of Datura Stramonium medicinal plant under stress allelopathic components of Eucalyptus camaldulensis. Journal of Rangeland. 12(4), 401-410. (In Persian with English abstract).
Sharma, P. and Dubey, R.S., 2005. Lead toxicity in plants. Brazilian Journal of Plant Physiology. 17(1), 35-52.
Soltani, A., 2001. Genetic variation for and interrelationships among seed vigor traits in wheat from the Caspian Sea coast of Iran. Seed Science and Technology. 29, 653-662.
Stefani, A., Arduini, I. and Onnis, A., 1991. Juncus acutus: germination and initial growth in presence of heavy metals. Annales Botanici Fennici. 28(1), 37-43.
Tatian, M.R., Tamartash, R., Heshmati, S. and Saeedi Garaghani, H.R., 2014. A study of Secale montanum seed response to stress caused by Pb and Cu pollutant elements. Journal of Natural Environment. 66(4), 389-397. (In Persian with English abstract).
Thounaojam, T.C., Panda, P., Mazumdar, P., Kumar, D., Sharma, G.D., Sahoo, L. and Sanjib, P., 2012. Excess copper induced oxidative stress and response of antioxidants in rice. Plant Physiology and Biochemistry. 53, 33-39.
Trofimova, T.A., Hossain, A. and da Silva, J.A.T., 2012. The ability of medical halophytes to phytoremediate soil contaminated by salt and heavy metals in Lower Volga, Russia. The Asian and Australasian Journal of Plant Science and Biotechnology. 6(1), 108-114.
Vogel-Mikus, K., Pongrac, P., Kump, P., Necemer, M., Simcic, J., Pelicon, P., Budnar, M., Povh, B. and Regvar, M., 2007. Localisation and quantification of elements within seeds of Cd/Zn hyperaccumulator Thlaspi praecox by micro-PIXE. Environmental Pollution. 147(1), 50-59.
Wani, P.A., Khan, M.S. and Zaidi, A., 2012. Toxic effects of heavy metal on germination and physiological processes of plants. In: Zaidi, A., Wani, P.A. and Khan, M.S. (Eds.), Toxicity of Heavy Metal to Legumes and Bioremediation. Springer, Verlag, Vienna, pp. 45-66.
Wierzbicka, M. and Obidzinska, J., 1998. The effect of lead on seed imbibition and germination in different plant species. Plant Science. 137(2), 155-171.